Identification of the cortical neurons that mediate antidepressant responses.

Our understanding of current treatments for depression, and the development of more specific therapies, is limited by the complexity of the circuits controlling mood and the distributed actions of antidepressants. Although the therapeutic efficacy of serotonin-specific reuptake inhibitors (SSRIs) is correlated with increases in cortical activity, the cell types crucial for their action remain unknown. Here we employ bacTRAP translational profiling to show that layer 5 corticostriatal pyramidal cells expressing p11 (S100a10) are strongly and specifically responsive to chronic antidepressant treatment. This response requires p11 and includes the specific induction of Htr4 expression. Cortex-specific deletion of p11 abolishes behavioral responses to SSRIs, but does not lead to increased depression-like behaviors. Our data identify corticostriatal projection neurons as critical for the response to antidepressants, and suggest that the regulation of serotonergic tone in this single cell type plays a pivotal role in antidepressant therapy.

Enhancer sequence variants and transcription-factor deregulation synergize to construct pathogenic regulatory circuits in B-cell lymphoma.

Most B-cell lymphomas arise in the germinal center (GC), where humoral immune responses evolve from potentially oncogenic cycles of mutation, proliferation, and clonal selection. Although lymphoma gene expression diverges significantly from GC B cells, underlying mechanisms that alter the activities of corresponding regulatory elements (REs) remain elusive. Here we define the complete pathogenic circuitry of human follicular lymphoma (FL), which activates or decommissions REs from normal GC B cells and commandeers enhancers from other lineages. Moreover, independent sets of transcription factors, whose expression was deregulated in FL, targeted commandeered versus decommissioned REs. Our approach revealed two distinct subtypes of low-grade FL, whose pathogenic circuitries resembled GC B or activated B cells. FL-altered enhancers also were enriched for sequence variants, including somatic mutations, which disrupt transcription-factor binding and expression of circuit-linked genes. Thus, the pathogenic regulatory circuitry of FL reveals distinct genetic and epigenetic etiologies for GC B-cell transformation.

Children diagnosed with presymptomatic type 1 diabetes through public health screening have milder diabetes at clinical manifestation.

AIMS/HYPOTHESIS: We aimed to determine whether disease severity was reduced at onset of clinical (stage 3) type 1 diabetes in children previously diagnosed with presymptomatic type 1 diabetes in a population-based screening programme for islet autoantibodies. METHODS: Clinical data obtained at diagnosis of stage 3 type 1 diabetes were evaluated in 128 children previously diagnosed with presymptomatic early-stage type 1 diabetes between 2015 and 2022 in the Fr1da study and compared with data from 736 children diagnosed with incident type 1 diabetes between 2009 and 2018 at a similar age in the DiMelli study without prior screening. RESULTS: At the diagnosis of stage 3 type 1 diabetes, children with a prior early-stage diagnosis had lower median HbA1c (51 mmol/mol vs 91 mmol/mol [6.8% vs 10.5%], p<0.001), lower median fasting glucose (5.3 mmol/l vs 7.2 mmol/l, p<0.05) and higher median fasting C-peptide (0.21 nmol/l vs 0.10 nmol/l, p<0.001) compared with children without previous early-stage diagnosis. Fewer participants with prior early-stage diagnosis had ketonuria (22.2% vs 78.4%, p<0.001) or required insulin treatment (72.3% vs 98.1%, p<0.05) and only 2.5% presented with diabetic ketoacidosis at diagnosis of stage 3 type 1 diabetes. Outcomes in children with a prior early-stage diagnosis were not associated with a family history of type 1 diabetes or diagnosis during the COVID-19 pandemic. A milder clinical presentation was observed in children who participated in education and monitoring after early-stage diagnosis. CONCLUSIONS/INTERPRETATION: Diagnosis of presymptomatic type 1 diabetes in children followed by education and monitoring improved clinical presentation at the onset of stage 3 type 1 diabetes.

Ribosome-binding and anti-microbial studies of the mycinamicins, 16-membered macrolide antibiotics from Micromonospora griseorubida.

Macrolides have been effective clinical antibiotics for over 70 years. They inhibit protein biosynthesis in bacterial pathogens by narrowing the nascent protein exit tunnel in the ribosome. The macrolide class of natural products consist of a macrolactone ring linked to one or more sugar molecules. Most of the macrolides used currently are semi-synthetic erythromycin derivatives, composed of a 14- or 15-membered macrolactone ring. Rapidly emerging resistance in bacterial pathogens is among the most urgent global health challenges, which render many antibiotics ineffective, including next-generation macrolides. To address this threat and advance a longer-term plan for developing new antibiotics, we demonstrate how 16-membered macrolides overcome erythromycin resistance in clinically isolated Staphylococcus aureus strains. By determining the structures of complexes of the large ribosomal subunit of Deinococcus radiodurans (D50S) with these 16-membered selected macrolides, and performing anti-microbial studies, we identified resistance mechanisms they may overcome. This new information provides important insights toward the rational design of therapeutics that are effective against drug resistant human pathogens.

Cancer Education for High School Students in the Northwest Arctic Increases Knowledge and Inspires Intent to Share Information and Reduce Cancer Risk.

Culturally appropriate cancer education is an opportunity to reduce health inequities in cancer. This manuscript describes the outcomes of piloting cancer education for youth in the Northwest Arctic region of Alaska. The project began due to community concerns, was focused through sharing circles conducted in the region, and was guided by a community advisory board. The project was based on the principles of Community Based Participatory Action Research (CBPAR), honored Indigenous Ways of Knowing, and was grounded in Empowerment Theory. In response to community requests, eleven cancer education lessons were developed for young people in the Northwest Arctic. Several lessons were piloted in spring 2022. Each participant was invited to complete a pre-lesson and a post-lesson survey. A total of 113 surveys were completed from five different lessons: 66 pre-lesson surveys and 47 post-lesson surveys. Respondents' mean cancer knowledge scores were significantly higher after the Cancer Basics lesson. On 98% of post-lesson surveys, respondents said they planned to share cancer education messages such as staying tobacco-free and increasing physical activity with others, including their family, friends, and community members. On 93% of the post-lesson surveys, respondents indicated they planned to make changes to reduce their own personal cancer risk, including by staying tobacco-free, eating healthier, and increasing physical activity. "Cancer is serious, and something we should start talking about".

Mercury Concentrations in Whale Shark (Rhincodon typus) Embryo Muscle Tissue.

Mercury (Hg) is known to be maternally transferred during embryonic development in sharks; however, Hg concentrations in embryos of filter feeding shark species have not previously been reported. This study measured the total Hg (THg) concentration in muscle tissue of 27 embryos taken from a pregnant whale shark (Rhincodon typus) landed in Taiwan in 1995 and the mean THg concentration compared to the mean muscle THg concentration in embryos from other shark species. The mean (± standard deviation) THg concentration in whale shark embryos was 0.0762 ± 0.0163 µg/g dry weight (0.0224 ± 0.0054 µg/g wet weight). There was no relationship between muscle THg concentration and body length and no significant difference in THg concentration between male and female embryos (p > 0.05). Whale shark embryos have the lowest reported muscle THg concentrations compared to literature values for muscle THg concentrations for embryos from other shark species.

Association of Race and Neighborhood Disadvantage with Patient Engagement in a Home-Based COVID-19 Remote Monitoring Program.

BACKGROUND: COVID-positive outpatients may benefit from remote monitoring, but such a program often relies on smartphone apps. This may introduce racial and socio-economic barriers to participation. Offering multiple methods for participation may address these barriers. OBJECTIVES: (1) To examine associations of race and neighborhood disadvantage with patient retention in a monitoring program offering two participation methods. (2) To measure the association of the program with emergency department visits and hospital admissions. DESIGN: Retrospective propensity-matched cohort study. PARTICIPANTS: COVID-positive outpatients at a single university-affiliated healthcare system and propensity-matched controls. INTERVENTIONS: A home monitoring program providing daily symptom tracking via patient portal app or telephone calls. MAIN MEASURES: Among program enrollees, retention (until 14 days, symptom resolution, or hospital admission) by race and neighborhood disadvantage, with stratification by program arm. In enrollees versus matched controls, emergency department utilization and hospital admission within 30 days. KEY RESULTS: There were 7592 enrolled patients and 9710 matched controls. Black enrollees chose the telephone arm more frequently than White enrollees (68% versus 44%, p = 0.009), as did those from more versus less disadvantaged neighborhoods (59% versus 43%, p = 0.02). Retention was similar in Black enrollees and White enrollees (63% versus 62%, p = 0.76) and in more versus less disadvantaged neighborhoods (63% versus 62%, p = 0.44). When stratified by program arm, Black enrollees had lower retention than White enrollees in the app arm (49% versus 55%, p = 0.01), but not in the telephone arm (69% versus 71%, p = 0.12). Compared to controls, enrollees more frequently visited the emergency department (HR 1.71 [95% CI 1.56-1.87]) and were admitted to the hospital (HR 1.16 [95% CI 1.02-1.31]). CONCLUSIONS: In a COVID-19 remote patient monitoring program, Black enrollees preferentially selected, and had higher retention in, telephone- over app-based monitoring. As a result, overall retention was similar between races. Remote monitoring programs with multiple modes may reduce barriers to participation.

Primary Care Practice Transformation in the Era of COVID-19 and Beyond: Key Principles for General Internal Medicine Practitioners.

As members of the Clinical Practice Committee (CPC) of the Society for General Internal Medicine (SGIM), we support practice innovation and transformation to achieve a more just system by which all people can achieve and maintain optimal health. The COVID-19 pandemic has tested the US healthcare delivery system and sharpened our national awareness of long-standing and ingrained system shortcomings. In the face of crisis, SGIM members innovated and energetically mobilized to focus on the immediate needs of our patients and communities. Reflecting on these experiences, we are called to consider what was learned from the pandemic that applies to the future of healthcare delivery. CPC members include leaders in primary care delivery, practice finance, quality of care, patient safety, hospital practice, and health policy. CPC members provide expertise in clinical practice, serving as primary care doctors, hospitalists, and patient advocates who understand the intensity of care needed for those with severe COVID-19 infections, the disproportionate impact of the pandemic on Black and Brown communities, the struggles created for those with poor access to care, and the physical and emotional impact it has placed on patients, families, and clinicians. In this consensus statement, we summarize lessons learned from the 2020-2021 pandemic and their broader implications for reform in healthcare delivery. We provide a platform for future work by identifying many interactive elements of healthcare delivery that must be simultaneously addressed in order to ensure that care is accessible, equitably provided, patient-centered, and cost-effective.

Serotonin receptor 4 in the hippocampus modulates mood and anxiety.

Serotonin receptor 4 (5-HT4R) plays an important role in regulating mood, anxiety, and cognition, and drugs that activate this receptor have fast-acting antidepressant (AD)-like effects in preclinical models. However, 5-HT4R is widely expressed throughout the central nervous system (CNS) and periphery, making it difficult to pinpoint the cell types and circuits underlying its effects. Therefore, we generated a Cre-dependent 5-HT4R knockout mouse line to dissect the function of 5-HT4R in specific brain regions and cell types. We show that the loss of functional 5-HT4R specifically from excitatory neurons of hippocampus led to robust AD-like behavioral responses and an elevation in baseline anxiety. 5-HT4R was necessary to maintain the proper excitability of dentate gyrus (DG) granule cells and cell type-specific molecular profiling revealed a dysregulation of genes necessary for normal neural function and plasticity in cells lacking 5-HT4R. These adaptations were accompanied by an increase in the number of immature neurons in ventral, but not dorsal, dentate gyrus, indicating a broad impact of 5-HT4R loss on the local cellular environment. This study is the first to use conditional genetic targeting to demonstrate a direct role for hippocampal 5-HT4R signaling in modulating mood and anxiety. Our findings also underscore the need for cell type-based approaches to elucidate the complex action of neuromodulatory systems on distinct neural circuits.

Chewing gum reduces visually induced motion sickness.

Visually induced motion sickness (VIMS) is a common side-effect of exposure to virtual reality (VR). Its unpleasant symptoms may limit the acceptance of VR technologies for training or clinical purposes. Mechanical stimulation of the mastoid and diverting attention to pleasant stimuli-like odors or music have been found to ameliorate VIMS. Chewing gum combines both in an easy-to-administer fashion and should thus be an effective countermeasure against VIMS. Our study investigated whether gustatory-motor stimulation by chewing gum leads to a reduction of VIMS symptoms. 77 subjects were assigned to three experimental groups (control, peppermint gum, and ginger gum) and completed a 15-min virtual helicopter flight, using a VR head-mounted display. Before and after VR exposure, we assessed VIMS with the Simulator Sickness Questionnaire (SSQ), and during the virtual flight once every minute with the Fast Motion Sickness Scale (FMS). Chewing gum (peppermint gum: M = 2.44, SD = 2.67; ginger gum: M = 2.57, SD = 3.30) reduced the peak FMS scores by 2.05 (SE = 0.76) points as compared with the control group (M = 4.56, SD = 3.52), p < 0.01, d = 0.65. Additionally, taste ratings correlated slightly negatively with both the SSQ and the peak FMS scores, suggesting that pleasant taste of the chewing gum is associated with less VIMS. Thus, chewing gum may be useful as an affordable, accepted, and easy-to-access way to mitigate VIMS in numerous applications like education or training. Possible mechanisms behind the effect are discussed.

Contemporary evolution rivals the effects of rhizobium presence on community and ecosystem properties in experimental mesocosms.

Because genotypes within a species commonly differ in traits that influence other species, whole communities, or even ecosystem functions, evolutionary change within one key species may affect the community and ecosystem processes. Here we use experimental mesocosms to test how the evolution of reduced cooperation in rhizobium mutualists in response to 20 years of nitrogen fertilization compares to the effects of rhizobium presence on soil nitrogen availability and plant community composition and diversity. The evolution of reduced rhizobium cooperation caused reductions in soil nitrogen, biological nitrogen fixation, and leaf nitrogen concentrations that were as strong as, or even stronger than, experimental rhizobium inoculation (presence/absence) treatments. Effects of both rhizobium evolution and rhizobium inoculation on legume dominance, plant community composition, and plant species diversity were often smaller in magnitude, but suggest that rhizobium evolution can alter the relative abundance of plant functional groups. Our findings indicate that the consequences of rapid microbial evolution for ecosystems and communities can rival the effects resulting from the presence or abundance of keystone mutualists.

Learning Together: Sharing Circles in Rural Alaska on Cancer Education Priorities for Youth.

Culturally relevant health promotion with youth is an opportunity to reduce health inequities in cancer. This manuscript describes sharing circles conducted with three communities in the Northwest Arctic region of Alaska. The circles were designed to begin understanding community priorities and lay the foundation to develop culturally relevant cancer education. The project was guided by the principles of Community-Based Participatory Action Research (CBPAR), honored Indigenous ways of knowing, and was grounded in Empowerment Theory. The project team facilitated 13 sharing circles in November 2019 in three communities in the Northwest Arctic. There were a total of 122 participants, including teachers/school staff (31%), community members (30%), high school students (23%), and health professionals (16%). The circles explored youth knowledge, perceptions, questions, concerns, and hopes for cancer information; community members' desires for youth knowledge about cancer; and how teachers would like content to be formatted for effective inclusion in their classrooms. Common themes from the sharing circles included a desire for information on cancer prevention (all 13 sharing circles) and a need for information on cancer risk factors (12). In most sharing circles, participants shared that cancer information for youth should include stories like those of local people, cancer survivors, and role models (11), visuals (8), and local data and statistics (8). In addition, teachers and school staff in all communities wanted an online resource for teaching about cancer in their classrooms that had short videos/visuals with related lesson plans and activities."If I learn, I can reduce the chance of getting cancer in the future."

The Influence of Physiological and Psychological Learning Mechanisms in Neurofeedback vs. Mental Imagery Against Binge Eating.

In biofeedback research, the debate on physiological versus psychological learning has a long tradition and is still relevant today, regarding new developments of biofeedback for behavior modification. Analyzing the role of these learning mechanisms may help improving the protocols and answer the question, whether feedback of physiological functions is necessary to modify a target behavior. We explored the presence and impact of physiological (EEG changes) versus psychological learning (changes in somatic self-efficacy) in a recently developed EEG neurofeedback protocol for binge eating. The protocol targets a reduction of food-cue induced cortical arousal through regulation of EEG high beta activity. In an experimental study accompanying a randomized controlled trial, pre and post treatment EEG measurements were analyzed in a neurofeedback group (n = 18) and an active mental imagery control group without physiological feedback (n = 18). Physiological learning in terms of EEG high beta reduction only occurred in the neurofeedback group. Post treatment, participants with successfully reduced binge eating episodes (≥ 50% reduction) showed lower EEG high beta activity than unsuccessful participants (p = .02) after neurofeedback, but not after mental imagery. Further, lower EEG high beta activity at post-treatment predicted fewer binge eating episodes in neurofeedback only. In mental imagery, somatic self-efficacy predicted treatment success instead of EEG activity. Altogether, the results indicate that physiological changes serve as a specific treatment mechanism in neurofeedback against binge eating. Reducing cortical arousal may improve eating behaviors and corresponding neurofeedback techniques should therefore be considered in future treatments.

FoxN3 is necessary for the development of the interatrial septum, the ventricular trabeculae and the muscles at the head/trunk interface in the African clawed frog, Xenopus laevis (Lissamphibia: Anura: Pipidae).

BACKGROUND: Fox genes are a large family of transcription factors that play diverse roles in the immune system, metabolism, cancer, cell cycle, and animal development. It has been shown that FoxN3 is indispensable for normal craniofacial development in the mouse and the African clawed frog, Xenopus laevis. Morpholino-mediated knockdown of FoxN3 in X. laevis delays overall development of early tadpole stages and causes eye defects, the absence of some cranial nerve branches, and malformations of the cranial skeleton and some cranial muscles, while the skeleton, nerves and muscles of the trunk are unaffected. RESULTS: We report a delay in heart morphogenesis, the absence of the interatrial septum, and a reduction and compaction of the ventricular trabeculation after knockdown of FoxN3 in X. laevis. Furthermore, we found malformations of the cucullaris and diaphragmatico-branchialis muscles, two head muscles that develop in the head/trunk interface of X. laevis. CONCLUSIONS: FoxN3 is necessary for the development of the interatrial septum and trabeculae in the frog heart, as well as the cranial muscles developing in the head/trunk interface. This gives the first evidence for a dependence on the head myogenic program of the cucullaris muscle in an anuran species.

Probing Selectivity and Creating Structural Diversity Through Hybrid Polyketide Synthases.

Engineering polyketide synthases (PKS) to produce new metabolites requires an understanding of catalytic points of failure during substrate processing. Growing evidence indicates the thioesterase (TE) domain as a significant bottleneck within engineered PKS systems. We created a series of hybrid PKS modules bearing exchanged TE domains from heterologous pathways and challenged them with both native and non-native polyketide substrates. Reactions pairing wildtype PKS modules with non-native substrates primarily resulted in poor conversions to anticipated macrolactones. Likewise, product formation with native substrates and hybrid PKS modules bearing non-cognate TE domains was severely reduced. In contrast, non-native substrates were converted by most hybrid modules containing a substrate compatible TE, directly implicating this domain as the major catalytic gatekeeper and highlighting its value as a target for protein engineering to improve analog production in PKS pathways.

Axonal ribosomes and mRNAs associate with fragile X granules in adult rodent and human brains.

Local mRNA translation in growing axons allows for rapid and precise regulation of protein expression in response to extrinsic stimuli. However, the role of local translation in mature CNS axons is unknown. Such a mechanism requires the presence of translational machinery and associated mRNAs in circuit-integrated brain axons. Here we use a combination of genetic, quantitative imaging and super-resolution microscopy approaches to show that mature axons in the mammalian brain contain ribosomes, the translational regulator FMRP and a subset of FMRP mRNA targets. This axonal translational machinery is associated with Fragile X granules (FXGs), which are restricted to axons in a stereotyped subset of brain circuits. FXGs and associated axonal translational machinery are present in hippocampus in humans as old as 57 years. This FXG-associated axonal translational machinery is present in adult rats, even when adult neurogenesis is blocked. In contrast, in mouse this machinery is only observed in juvenile hippocampal axons. This differential developmental expression was specific to the hippocampus, as both mice and rats exhibit FXGs in mature axons in the adult olfactory system. Experiments in Fmr1 null mice show that FMRP regulates axonal protein expression but is not required for axonal transport of ribosomes or its target mRNAs. Axonal translational machinery is thus a feature of adult CNS neurons. Regulation of this machinery by FMRP could support complex behaviours in humans throughout life.

Effects of Agricultural Management on Rhizosphere Microbial Structure and Function in Processing Tomato Plants.

Agricultural management practices affect bulk soil microbial communities and the functions they carry out, but it remains unclear how these effects extend to the rhizosphere in different agroecosystem contexts. Given close linkages between rhizosphere processes and plant nutrition and productivity, understanding how management practices impact this critical zone is of great importance to optimize plant-soil interactions for agricultural sustainability. A comparison of six paired conventional-organic processing tomato farms was conducted to investigate relationships between management, soil physicochemical parameters, and rhizosphere microbial community composition and functions. Organically managed fields were higher in soil total N and NO3-N, total and labile C, plant Ca, S, and Cu, and other essential nutrients, while soil pH was higher in conventionally managed fields. Differential abundance, indicator species, and random forest analyses of rhizosphere communities revealed compositional differences between organic and conventional systems and identified management-specific microbial taxa. Phylogeny-based trait prediction showed that these differences translated into more abundant pathogenesis-related gene functions in conventional systems. Structural equation modeling revealed a greater effect of soil biological communities than physicochemical parameters on plant outcomes. These results highlight the importance of rhizosphere-specific studies, as plant selection likely interacts with management in regulating microbial communities and functions that impact agricultural productivity.IMPORTANCE Agriculture relies, in part, on close linkages between plants and the microorganisms that live in association with plant roots. These rhizosphere bacteria and fungi are distinct from microbial communities found in the rest of the soil and are even more important to plant nutrient uptake and health. Evidence from field studies shows that agricultural management practices such as fertilization and tillage shape microbial communities in bulk soil, but little is known about how these practices affect the rhizosphere. We investigated how agricultural management affects plant-soil-microbe interactions by comparing soil physical and chemical properties, plant nutrients, and rhizosphere microbial communities from paired fields under organic and conventional management. Our results show that human management effects extend even to microorganisms living in close association with plant roots and highlight the importance of these bacteria and fungi to crop nutrition and productivity.

MicroFEWs: A Food-Energy-Water Systems Approach to Renewable Energy Decisions in Islanded Microgrid Communities in Rural Alaska.

In recent years, there has been increased recognition of the importance of a nexus approach to optimize food, energy, and water (FEW) security at regional and global scales. Remote communities in the Arctic and Subarctic regions in Alaska provide unique examples of closed and isolated systems, wherein the FEW nexus not only needs to be examined to lend resilience to these vulnerable communities but that could also serve as small-scale test beds for a wider and systematic understanding of the FEW nexus. In this short communication, looking at the FEW nexus in Cordova, Alaska, through an energy lens, we introduce an approach (referred to as the "MicroFEWs approach") that may assist remote communities in Alaska in making informed decisions regarding the use of renewable energy to increase FEW security. Our example uses the MicroFEWs approach to assess the impacts of increased renewable energy generation on FEW security in the community, more specifically to food security through potential changes to the community's fish processing industry. This approach can serve as a basis for investigating the FEW nexus in varying contexts and locales.

p11 and its role in depression and therapeutic responses to antidepressants.

Studies of the multifunctional protein p11 (also known as S100A10) are shedding light on the molecular and cellular mechanisms underlying depression. Here, we review data implicating p11 in both the amplification of serotonergic signalling and the regulation of gene transcription. We summarize studies demonstrating that levels of p11 are regulated in depression and by antidepressant regimens and, conversely, that p11 regulates depression-like behaviours and/or responses to antidepressants. Current and future studies of p11 may provide a molecular and cellular framework for the development of novel antidepressant therapies.

Unveiling sequential late-stage methyltransferase reactions in the meleagrin/oxaline biosynthetic pathway.

Antimicrobial and anti-proliferative meleagrin and oxaline are roquefortine C-derived alkaloids produced by fungi of the genus Penicillium. Tandem O-methylations complete the biosynthesis of oxaline from glandicoline B through meleagrin. Currently, little is known about the role of these methylation patterns in the bioactivity profile of meleagrin and oxaline. To establish the structural and mechanistic basis of methylation in these pathways, crystal structures were determined for two late-stage methyltransferases in the oxaline and meleagrin gene clusters from Penicillium oxalicum and Penicillium chrysogenum. The homologous enzymes OxaG and RoqN were shown to catalyze penultimate hydroxylamine O-methylation to generate meleagrin in vitro. Crystal structures of these enzymes in the presence of methyl donor S-adenosylmethionine revealed an open active site, which lacks an apparent base indicating that catalysis is driven by proximity effects. OxaC was shown to methylate meleagrin to form oxaline in vitro, the terminal pathway product. Crystal structures of OxaC in a pseudo-Michaelis complex containing sinefungin and meleagrin, and in a product complex containing S-adenosyl-homocysteine and oxaline, reveal key active site residues with His313 serving as a base that is activated by Glu369. These data provide structural insights into the enzymatic methylation of these alkaloids that include a rare hydroxylamine oxygen acceptor, and can be used to guide future efforts towards selective derivatization and structural diversification and establishing the role of methylation in bioactivity.